Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies.

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Abstract Text:

J Ronco,A Charbit,M Hofnung,

LamB, an integral outer membrane protein of E coli K12, is highly resistant to protease digestion. We had previously genetically inserted a foreign sequence corresponding to an epitope from the poliovirus next to amino acids 146, 153, 189, and 374 of LamB. In 3 cases (sites 146, 153, 374), insertion of the foreign peptide did not extensively affect the functions of LamB (and therefore folding). In 2 cases (sites 146 and 374) the polio virus epitope was detectable on the bacterial surface with a specific monoclonal antibody. We show here that the 4 modified proteins are sensitive to trypsin, including on intact cells. The sizes of the major cleavage products is that expected for proteolysis at or near the sequences inserted. In 1 case (site 153), this was directly demonstrated by protein sequencing. The results confirm the cell surface exposure of the regions of residues 153 and 374 and provide information on the regions around residues 146 and 189. Perspectives and limitations of this approach for fine studies on the mode of insertion of membrane proteins are briefly discussed.

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Publishing Authors By Initials

J Ronco,A Charbit,M Hofnung,

For similar proteins: viral proteins research abstracts see: proteins: viral proteins research

PUBMED ID PMID:

MEDLINE DATE: 1990 Feb-Mar

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Biochimie

VOLUME: 72

Page Numbers: 183-9

Journal Abbreviation: Biochimie

ISSN: 0300-9084

DAY: 27

MONTH: 11

YEAR: 2007

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 1264604

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Keywords Mesh Terms:

KEYWORDS: Viral Proteins

MESH TERMS: genetics

Chemical & Substance for Abstract: Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies. Information

Substance Name: Trypsin

Registry Number: EC 3.4.21.4

Grant and Affiliation Information for Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies.

AFFILIATION: Unité de Programmation Moléculaire et Toxicologie Génétique, CNRS UA271 INSERM U163, Institut Pasteur, Paris, France.

Country: FRANCE

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: Biochimie

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies Related Publications

• A genetic system to elicit and monitor antipeptide antibodies without peptide synthesis.
• A system for genetic analysis in gene lamB: first results with lambda-resistant tight mutants.
• An intelligent channel (and more).
• Antibodies against synthetic peptides and the topology of LamB, an outer membrane protein from Escherichia coli K12.
• Bacteriophage lambda receptor site on the Escherichia coli K-12 LamB protein.
• Creation of targets for proteolytic cleavage in the LamB protein of E coli K12 by genetic insertion of foreign sequences: implications for topological studies.
• DNA sequence analysis of the lamB gene from Klebsiella pneumoniae: implications for the topology and the pore functions in maltoporin.
• Expression and immunogenicity of the V3 loop from the envelope of human immunodeficiency virus type 1 in an attenuated aroA strain of Salmonella typhimurium upon genetic coupling to two Escherichia coli carrier proteins.
• Expression of a poliovirus neutralization epitope at the surface of recombinant bacteria: first immunization results.
• Expression of foreign antigenic determinants on bacterial envelope proteins.
• Further sequence analysis of the phage lambda receptor site. Possible implications for the organization of the lamB protein in Escherichia coli K12.
• Gene sequence of the lambda receptor, an outer membrane protein of E. coli K12.
• Genetic study of a membrane protein: DNA sequence alterations due to 17 lamB point mutations affecting adsorption of phage lambda.
• Immunodominance of a recombinant T-cell epitope depends on its molecular environment.
• Immunogenicity and antigenicity of conserved peptides from the envelope of HIV-1 expressed at the surface of recombinant bacteria.
• Immunogenicity of viral B- and T-cell epitopes expressed in recombinant bacterial proteins.
• In situ enzyme immunodetection of surface or intracellular bacterial antigens using nitrocellulose sheets.
• In vivo and in vitro studies of transmembrane beta-strand deletion, insertion or substitution mutants of the Escherichia coli K-12 maltoporin.
• Isolation of different bacteriophages using the LamB protein for adsorption on Escherichia coli K-12.
• Linker mutagenesis in the gene of an outer membrane protein of Escherichia coli, lamB.
• Maltose transport and starch binding in phage-resistant point mutants of maltoporin. Functional and topological implications.
• Mutagenesis by random linker insertion into the lamB gene of Escherichia coli K12.
• Mutations in gene lamB: studies on structure and topology of an E. coli outer membrane protein.
• Negative dominance in gene lamB: random assembly of secreted subunits issued from different polysomes.
• Permissive sites and topology of an outer membrane protein with a reporter epitope.
• Presentation of two epitopes of the preS2 region of hepatitis B virus on live recombinant bacteria.
• The cellular location of a foreign B cell epitope expressed by recombinant bacteria determines its T cell-independent or T cell-dependent characteristics.
• The maltoporin of Salmonella typhimurium: sequence and folding model.
• [DNA sequence encoding the signal peptide of the lambda receptor in E. coli K 12]
• rho Mutations restore lamB expression in E. coli K12 strains with an inactive malB region.